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nf-core/nascent

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Nextflow run with conda run with docker run with singularity Launch on Nextflow Tower

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Introduction

nf-core/nascent is a bioinformatics best-practice analysis pipeline for nascent transcript (NT) and Transcriptional Start Site (TSS) assays.

The pipeline is built using Nextflow, a workflow tool to run tasks across multiple compute infrastructures in a very portable manner. It uses Docker/Singularity containers making installation trivial and results highly reproducible. The Nextflow DSL2 implementation of this pipeline uses one container per process which makes it much easier to maintain and update software dependencies. Where possible, these processes have been submitted to and installed from nf-core/modules in order to make them available to all nf-core pipelines, and to everyone within the Nextflow community!

On release, automated continuous integration tests run the pipeline on a full-sized dataset on the AWS cloud infrastructure. This ensures that the pipeline runs on AWS, has sensible resource allocation defaults set to run on real-world datasets, and permits the persistent storage of results to benchmark between pipeline releases and other analysis sources.The results obtained from the full-sized test can be viewed on the nf-core website.

Pipeline summary

  1. Read QC (FastQC)
  2. Adapter and quality trimming (fastp)
  3. Alignment
    1. bwa
    2. bwamem2
    3. DRAGMAP
  4. Sort and index alignments (SAMtools)
  5. UMI-based deduplication (UMI-tools)
  6. Duplicate read marking (picard MarkDuplicates)
  7. Quality Control
    1. RSeQC - Various RNA-seq QC metrics
    2. Preseq - Estimation of library complexity
    3. BBMap - Analyzes the sequencing coverage
  8. Coverage Graphs
    1. Create bedGraph coverage files (BEDTools
    2. Create bigWig coverage files (deeptools)
  9. Transcript identification
    1. HOMER
    2. GroHMM
    3. PINTS
  10. Quantification of Genes and Nascent Transcripts (featureCounts)
  11. Aggregate report describing results and QC from the whole pipeline (MultiQC)

Usage

Note

If you are new to Nextflow and nf-core, please refer to this page on how to set-up Nextflow. Make sure to test your setup with -profile test before running the workflow on actual data.

Now, you can run the pipeline using:

nextflow run nf-core/nascent \
   -profile <docker/singularity/.../institute> \
   --input samplesheet.csv \
   --outdir <OUTDIR>

Warning

Please provide pipeline parameters via the CLI or Nextflow -params-file option. Custom config files including those provided by the -c Nextflow option can be used to provide any configuration except for parameters; see docs.

For more details and further functionality, please refer to the usage documentation and the parameter documentation.

Pipeline output

To see the results of an example test run with a full size dataset refer to the results tab on the nf-core website pipeline page. For more details about the output files and reports, please refer to the output documentation.

Credits

nf-core/nascent was originally written by Ignacio Tripodi (@ignaciot) and Margaret Gruca (@magruca).

The pipeline was re-written in Nextflow DSL2 by Edmund Miller (@Emiller88) and Sruthi Suresh (@sruthipsuresh) from The Functional Genomics Laboratory at The Univeristy of Texas at Dallas

We thank the following people for their extensive assistance in the development of this pipeline:

Contributions and Support

If you would like to contribute to this pipeline, please see the contributing guidelines.

For further information or help, don't hesitate to get in touch on the Slack #nascent channel (you can join with this invite).

Citations

If you use nf-core/nascent for your analysis, please cite it using the following doi: 10.5281/zenodo.7245273

An extensive list of references for the tools used by the pipeline can be found in the CITATIONS.md file.

You can cite the nf-core publication as follows:

The nf-core framework for community-curated bioinformatics pipelines.

Philip Ewels, Alexander Peltzer, Sven Fillinger, Harshil Patel, Johannes Alneberg, Andreas Wilm, Maxime Ulysse Garcia, Paolo Di Tommaso & Sven Nahnsen.

Nat Biotechnol. 2020 Feb 13. doi: 10.1038/s41587-020-0439-x.

nascent's People

Contributors

apeltzer avatar drpatelh avatar edmundmiller avatar ignaciot avatar kevinmenden avatar nf-core-bot avatar nithyaj12 avatar sruthipsuresh avatar

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nascent's Issues

"ERROR ~ fromIndex = -1" during samtools sort step

Description of the bug

I have tried running the pipeline with both bwa and bwamem, either way everything seems to run fine up until the samtools sort step, which does appear to complete on the first sample, but not the second sample regardless of which sample is second. I have tried running samtools sort on the bams created by the pipeline independently and it works just fine for all of my samples when I run it. The error message I am given from the pipeline is: "ERROR ~ fromIndex = -1"

Command used and terminal output

NXF_VER=23.04.1 /config/binaries/nextflow/23.04.1/nextflow run nf-core/nascent -profile singularity \
    --input /scratch/teams/dawson_genomics/Projects/MYC/230427_PROseq/scripts/samplesheet.csv \
    --outdir /scratch/teams/dawson_genomics/Projects/MYC/230427_PROseq/nf-core_nascent \
    --genome GRCh38 \
    --aligner bwamem2 \
    --multiqc_title nf-core_nacent_multiqc \
    --assay_type PROseq \
    -resume

ERROR ~ fromIndex = -1

 -- Check script '/home/agillespie/.nextflow/assets/nf-core/nascent/./workflows/nascent.nf' at line: 222 or see '.nextflow.log' file for more details

Relevant files

nextflow.log
samplesheet.csv

System information

Nextflow version: 23.04.1
Hardware: HPC
Executor: slurm
Container: singularity
OS: CentOS 7 Linux
nf-core/nascent v2.1.1-g9ff33c7
samtools version: 1.17

Error executing process > 'NFCORE_NASCENT:NASCENT:ALIGN_BWA:BWA_MEM (1)'

Description of the bug

When I ran the nfcore/ nascent pipeline for PRO-seq data using the default aligner in dev mode I ran into this issue.

I used 2 commands for 2 different runs, and both yielded the same Error message in the end.

Command used and terminal output

nextflow run nf-core/nascent -r dev --input samplesheet.csv --outdir nascentoutput --fasta hg38_noalt/fasta/genome_noalt.fa --gtf hg38_annotations/ucsc/hg38.ncbiRefSeq.gtf --bwa_index BWA_Human/bwa_hg38_noalt_index/ --assay_type PROseq --max_cpus 10 -profile singularity


nextflow run nf-core/nascent -r dev --input samplesheet.csv --outdir nascentoutput --genome hg38 --assay_type PROseq --max_cpus 10 -profile singularity

Error executing process > 'NFCORE_NASCENT:NASCENT:ALIGN_BWA:BWA_MEM (1)'
Caused by:
  Not a valid path value type: org.codehaus.groovy.runtime.NullObject (null)
Tip: view the complete command output by changing to the process work dir and entering the command `cat .command.out`

Relevant files

No response

System information

Nextflow Version: 21.10.6
Hardware Desktop
Executor Slurm
Container Engine Singularity
OS Linux

Which assay type (GRO-seq, PRO-seq ... etc) should I use for EU-seq?

Description of feature

My question is which assay type (GRO-seq, PRO-seq .. etc.,) should I use for EU-seq analysis? This is the outline of EU-seq (PMID: 34485932). Briefly, nascent RNA is labeled by 5-EU in vivo. Afterword, EU-labeled RNAs are enriched, and reverse transcribed and amplified cDNAs having barcodes are sequenced. It looks like GRO-seq or PRO-seq but a bit different because in EU-seq experiment nascent RNAs are labeled in vivo without nuclear isolation and cell lysis steps.

Fix GroHMM on full runs

Samplesheet:

sample,fastq_1,fastq_2
GM0h_REP1,../data/groseq_raw/GM0h.fastq.gz,

Data available here:

Error:

Caused by:
  Process `NFCORE_NASCENT:NASCENT:GROHMM:GROHMM_TRANSCRIPTCALLING (GM0h)` terminated with an error exit status (1)

Command executed:

  transcriptcalling_grohmm.R \
      --bam_file GM0h.sorted.bam \
       \
      --outprefix GM0h \
      --gtf genes.gtf \                                                                                          --outdir ./ \
      --cores 12 \


  cat <<-END_VERSIONS > versions.yml
  "NFCORE_NASCENT:NASCENT:GROHMM:GROHMM_TRANSCRIPTCALLING":
      r-base: $(echo $(R --version 2>&1) | sed 's/^.*R version //; s/ .*$//')
      bioconductor-grohmm: $(Rscript -e "library(groHMM); cat(as.character(packageVersion('groHMM')))")
  END_VERSIONS

Command exit status:
  1

Command output:
  (empty)

Command error:
  Error in Fp[[i]] + 1 : non-numeric argument to binary operator
  Calls: detectTranscripts
  In addition: Warning messages:
  1: In mclapply(readsList, function(x) { :
    all scheduled cores encountered errors in user code
  2: In mclapply(readsList, function(x) { :
    all scheduled cores encountered errors in user code
  Execution halted

Work dir:
  /petastore/ganymede/home/eam150030/nascent/work/f0/e794715b51ad2b1036c36c8fb7b410

Tip: view the complete command output by changing to the process work dir and entering the command `cat .command.out`

Many Pipeline running errors (iGenomes, hisat index, then bbduk error)

I've had many errors thrown when trying to run this pipeline (using docker). First, using igenome (e.g. --genome GRCh37) stops with this error: "No such variable: ch_fasta_for_hisat_index" (The output shows that it is using the correct S3 fasta file). I then added the --hisat_indices reference to downloaded hisat2 indices for grch37, but the pipeline still stopped with the same error. When I then changed the genome reference to --fasta and pointed to a GRCh37 fasta file, it ran, but stopped with this message:

Error executing process > 'bbduk ([redacted].fastq)'

Caused by:
Missing output file(s) *.trim.fastq expected by process bbduk ([redacted].fastq)

and

Set error histogram output to [redacted].ehist.txt
Exception in thread "main" java.lang.AssertionError: Can't find reference file /home/david/.nextflow/assets/nf-core/nascent/assets/adapters.fa
at jgi.BBDukF.(BBDukF.java:556)
p: wheat jgi.BBDukF.main(BBDukF.java:67)

The file it can't find is indeed present in the assets folder.

Finally, I tried adding the --saveAll flag, that didn't help. I'm now stuck. I'd really like to use this pipeline, I've used the RNAseq and chipseq nf-core pipelines and they are fantastic.

FastQC ran fine! The bbduk program is what's choking...

thanks

Clean up published files

Description of feature

I think there are a lot of extra files that are published currently, such as the bam files from alignment before they're dedupped.

Issues of creating BWA index with custom fasta and GTF

Description of the bug

I used custom fasta and GTF files to use nfcore nascent and got the following error messages.

Error executing process > 'NFCORE_NASCENT:NASCENT:PREPARE_GENOME:BWA_INDEX (null)'
Caused by:
Not a valid path value type: groovyx.gpars.dataflow.DataflowVariable (DataflowVariable(value=null))

The point is that this custom reference genome is working in nfcore RNA-seq analysis but not in nfcore nascent.
Edmund Miller advised me to use --genome GRCh38 instead of the custom reference genome and --genome GRCh38 actually worked.

Based on these observations, we would think that this is a bug of BWA index creation when we put custom reference genome on parameters.

Command used and terminal output

/home/yhigashi/nextflow run nf-core/nascent -r 2.1.1 -work-dir /home/yhigashi/tnfil4_euseq -params-file /home/yhigashi/tnfil4_euseq/230920_nf-params_v2.json -profile singularity --save_reference



N E X T F L O W  ~  version 22.10.5
Launching `https://github.com/nf-core/nascent` [distraught_newton] DSL2 - revision: 9ff33c7896 [2.1.1]


------------------------------------------------------
                                        ,--./,-.
        ___     __   __   __   ___     /,-._.--~'
  |\ | |__  __ /  ` /  \ |__) |__         }  {
  | \| |       \__, \__/ |  \ |___     \`-._,-`-,
                                        `._,._,'
  nf-core/nascent v2.1.1-g9ff33c7
------------------------------------------------------
Core Nextflow options
  revision       : 2.1.1
  runName        : distraught_newton
  containerEngine: singularity
  launchDir      : /rshare1/ZETTAI_path_WA_slash_home_KARA/home/yhigashi
  workDir        : /home/yhigashi/tnfil4_euseq
  projectDir     : /home/yhigashi/.nextflow/assets/nf-core/nascent
  userName       : yhigashi
  profile        : singularity
  configFiles    : /home/yhigashi/.nextflow/assets/nf-core/nascent/nextflow.config

Input/output options
  input          : /home/yhigashi/tnfil4_euseq/230920_samplesheet.csv
  outdir         : /home/yhigashi/nextflow_results/tnfil4_nextflow_euseq_results
  email          : [email protected]

Transcript Identification Options
  assay_type     : GROseq

Reference genome options
  fasta          : /home/yhigashi/Homo_sapiens.GRCh38.dna_sm.primary_assembly.fa.gz
  gtf            : /home/yhigashi/Homo_sapiens.GRCh38.110.gtf.gz
  save_reference : true

!! Only displaying parameters that differ from the pipeline defaults !!
------------------------------------------------------
If you use nf-core/nascent for your analysis please cite:

* The pipeline
  https://doi.org/10.5281/zenodo.7245273

* The nf-core framework
  https://doi.org/10.1038/s41587-020-0439-x

* Software dependencies
  https://github.com/nf-core/nascent/blob/master/CITATIONS.md
------------------------------------------------------
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[-        ] process > NFCORE_NASCENT:NASCENT:CUST... -
[-        ] process > NFCORE_NASCENT:NASCENT:MULTIQC -
Error executing process > 'NFCORE_NASCENT:NASCENT:PREPARE_GENOME:BWA_INDEX (null)'

Caused by:
  Not a valid path value type: groovyx.gpars.dataflow.DataflowVariable (DataflowVariable(value=null))


Tip: you can try to figure out what's wrong by changing to the process work dir and showing the script file named `.command.sh`



[-        ] process > NFCORE_NASCENT:NASCENT:PREP... -
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[-        ] process > NFCORE_NASCENT:NASCENT:ALIG... -
[-        ] process > NFCORE_NASCENT:NASCENT:ALIG... -
[-        ] process > NFCORE_NASCENT:NASCENT:ALIG... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:COVE... -
[-        ] process > NFCORE_NASCENT:NASCENT:COVE... -
[-        ] process > NFCORE_NASCENT:NASCENT:COVE... -
[-        ] process > NFCORE_NASCENT:NASCENT:COVE... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:BED2SAF -
[-        ] process > NFCORE_NASCENT:NASCENT:SUBR... -
[-        ] process > NFCORE_NASCENT:NASCENT:SUBR... -
[-        ] process > NFCORE_NASCENT:NASCENT:CUST... -
[-        ] process > NFCORE_NASCENT:NASCENT:MULTIQC -
Execution cancelled -- Finishing pending tasks before exit
Pulling Singularity image https://depot.galaxyproject.org/singularity/ubuntu:20.04 [cache /home/yhigashi/tnfil4_euseq/singularity/depot.galaxyproject.org-singularity-ubuntu-20.04.img]
Pulling Singularity image https://depot.galaxyproject.org/singularity/python:3.8.3 [cache /home/yhigashi/tnfil4_euseq/singularity/depot.galaxyproject.org-singularity-python-3.8.3.img]
WARN: Singularity cache directory has not been defined -- Remote image will be stored in the path: /home/yhigashi/tnfil4_euseq/singularity -- Use env variable NXF_SINGULARITY_CACHEDIR to specify a different location
Error executing process > 'NFCORE_NASCENT:NASCENT:PREPARE_GENOME:BWA_INDEX (null)'

Caused by:
  Not a valid path value type: groovyx.gpars.dataflow.DataflowVariable (DataflowVariable(value=null))


Tip: you can try to figure out what's wrong by changing to the process work dir and showing the script file named `.command.sh`



[-        ] process > NFCORE_NASCENT:NASCENT:PREP... -
[-        ] process > NFCORE_NASCENT:NASCENT:PREP... -
[-        ] process > NFCORE_NASCENT:NASCENT:PREP... -
[-        ] process > NFCORE_NASCENT:NASCENT:PREP... -
[-        ] process > NFCORE_NASCENT:NASCENT:PREP... -
[-        ] process > NFCORE_NASCENT:NASCENT:PREP... -
[-        ] process > NFCORE_NASCENT:NASCENT:INPU... -
[-        ] process > NFCORE_NASCENT:NASCENT:FASTQC  -
[-        ] process > NFCORE_NASCENT:NASCENT:FASTP   -
[-        ] process > NFCORE_NASCENT:NASCENT:ALIG... -
[-        ] process > NFCORE_NASCENT:NASCENT:ALIG... -
[-        ] process > NFCORE_NASCENT:NASCENT:ALIG... -
[-        ] process > NFCORE_NASCENT:NASCENT:ALIG... -
[-        ] process > NFCORE_NASCENT:NASCENT:ALIG... -
[-        ] process > NFCORE_NASCENT:NASCENT:ALIG... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:COVE... -
[-        ] process > NFCORE_NASCENT:NASCENT:COVE... -
[-        ] process > NFCORE_NASCENT:NASCENT:COVE... -
[-        ] process > NFCORE_NASCENT:NASCENT:COVE... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:BED2SAF -
[-        ] process > NFCORE_NASCENT:NASCENT:SUBR... -
[-        ] process > NFCORE_NASCENT:NASCENT:SUBR... -
[-        ] process > NFCORE_NASCENT:NASCENT:CUST... -
[-        ] process > NFCORE_NASCENT:NASCENT:MULTIQC -
Execution cancelled -- Finishing pending tasks before exit
Pulling Singularity image https://depot.galaxyproject.org/singularity/ubuntu:20.04 [cache /home/yhigashi/tnfil4_euseq/singularity/depot.galaxyproject.org-singularity-ubuntu-20.04.img]
Pulling Singularity image https://depot.galaxyproject.org/singularity/python:3.8.3 [cache /home/yhigashi/tnfil4_euseq/singularity/depot.galaxyproject.org-singularity-python-3.8.3.img]
-[nf-core/nascent] Sent summary e-mail to [email protected] (sendmail)-
-[nf-core/nascent] Pipeline completed with errors-
WARN: Singularity cache directory has not been defined -- Remote image will be stored in the path: /home/yhigashi/tnfil4_euseq/singularity -- Use env variable NXF_SINGULARITY_CACHEDIR to specify a different location
Error executing process > 'NFCORE_NASCENT:NASCENT:PREPARE_GENOME:BWA_INDEX (null)'

Caused by:
  Not a valid path value type: groovyx.gpars.dataflow.DataflowVariable (DataflowVariable(value=null))


Tip: you can try to figure out what's wrong by changing to the process work dir and showing the script file named `.command.sh`



[-        ] process > NFCORE_NASCENT:NASCENT:PREP... -
[-        ] process > NFCORE_NASCENT:NASCENT:PREP... -
[-        ] process > NFCORE_NASCENT:NASCENT:PREP... -
[-        ] process > NFCORE_NASCENT:NASCENT:PREP... -
[-        ] process > NFCORE_NASCENT:NASCENT:PREP... -
[-        ] process > NFCORE_NASCENT:NASCENT:PREP... -
[-        ] process > NFCORE_NASCENT:NASCENT:INPU... -
[-        ] process > NFCORE_NASCENT:NASCENT:FASTQC  -
[-        ] process > NFCORE_NASCENT:NASCENT:FASTP   -
[-        ] process > NFCORE_NASCENT:NASCENT:ALIG... -
[-        ] process > NFCORE_NASCENT:NASCENT:ALIG... -
[-        ] process > NFCORE_NASCENT:NASCENT:ALIG... -
[-        ] process > NFCORE_NASCENT:NASCENT:ALIG... -
[-        ] process > NFCORE_NASCENT:NASCENT:ALIG... -
[-        ] process > NFCORE_NASCENT:NASCENT:ALIG... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:QUAL... -
[-        ] process > NFCORE_NASCENT:NASCENT:COVE... -
[-        ] process > NFCORE_NASCENT:NASCENT:COVE... -
[-        ] process > NFCORE_NASCENT:NASCENT:COVE... -
[-        ] process > NFCORE_NASCENT:NASCENT:COVE... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:TRAN... -
[-        ] process > NFCORE_NASCENT:NASCENT:BED2SAF -
[-        ] process > NFCORE_NASCENT:NASCENT:SUBR... -
[-        ] process > NFCORE_NASCENT:NASCENT:SUBR... -
[-        ] process > NFCORE_NASCENT:NASCENT:CUST... -
[-        ] process > NFCORE_NASCENT:NASCENT:MULTIQC -
Execution cancelled -- Finishing pending tasks before exit
Pulling Singularity image https://depot.galaxyproject.org/singularity/ubuntu:20.04 [cache /home/yhigashi/tnfil4_euseq/singularity/depot.galaxyproject.org-singularity-ubuntu-20.04.img]
Pulling Singularity image https://depot.galaxyproject.org/singularity/python:3.8.3 [cache /home/yhigashi/tnfil4_euseq/singularity/depot.galaxyproject.org-singularity-python-3.8.3.img]
-[nf-core/nascent] Sent summary e-mail to [email protected] (sendmail)-
-[nf-core/nascent] Pipeline completed with errors-
WARN: Singularity cache directory has not been defined -- Remote image will be stored in the path: /home/yhigashi/tnfil4_euseq/singularity -- Use env variable NXF_SINGULARITY_CACHEDIR to specify a different location
Error executing process > 'NFCORE_NASCENT:NASCENT:PREPARE_GENOME:BWA_INDEX (null)'

Caused by:
  Not a valid path value type: groovyx.gpars.dataflow.DataflowVariable (DataflowVariable(value=null))


Tip: you can try to figure out what's wrong by changing to the process work dir and showing the script file named `.command.sh`

Relevant files

230920_nextflow.log

System information

  1. Nextflow version 22.10.5
  2. SuperComputer SHIROKANE
  3. AGE (Altair Grid Engine)
  4. Singularity
  5. RedHat Enterprise Linux 8 HPC (x86_64)
  6. nfcore/nascent 2.1.1

Investigate adapter trimming

Description of the bug

I haven't seen it done, so not sure if it would be beneficial or not.

Command used and terminal output

No response

Relevant files

No response

System information

No response

Bedtools sort output isn't deterministic

Description of the bug

Continues to have different md5sums. Probably not coming from bedtools sort, but that's the only output getting caught that's changing things.

Command used and terminal output

No response

Relevant files

No response

System information

No response

Document grohmm Tuning

Description of feature

Add to usage docs, pros and cons of tuning and the automation of it.

Support STAR

Description of feature

Already got feedback that this might handle splice sites in eRNAs better

Optimize grohmm parameter tuning and transcript calling

Right now, we're parameter tuning is calling detectTranscripts and then we redo the identification in the transcript calling script, and then do the downstream steps.

Not the end of the world, but definitely a bottleneck in the workflow.

Going to be a followup to #65

Refactor Prepare Genome

Description of feature

I want to follow something similar to Sarek's preparegenome and let the logic live in modules.config

Add PINTS Visualizer

Description of feature

Visualize TSS signals

pints_visualizer -b GROcap_hg38.uniq.bam \
    -e GROcap \
    --mapq-threshold 255 \
    --chromosome-start-with chr \
    -o GROcap_hg38 \
    --filters U13369 chrM EBV

test_full profile is broken

Description of the bug

-profile test_full is broken, so megatests are not working for nascent.
I believe it's mainly due to missing params assay_type, due to the error message:

Missing required parameter: --assay_type

Command used and terminal output

No response

Relevant files

No response

System information

No response

Set up AWS Full tests

Description of feature

Opening this as a follow-up to #66. There are two full test datasets, but not setup to run on AWS full tests.

Flipped bigwig strands in PROseq samples

Description of the bug

Hi Edmund,
I am raising a ticket here for the strand-flipping issue I mentioned on slack.

Bug:
BigWigs generated for PROseq samples appear to be flipped. The strand carrying the gene body signal is inconsistent with gene direction.
Example: GUSB gene on the negative strand. Genebody signal in the public Jurkat sample processed with the v2.1.1 pipeline is on the positive strand (UCSC Browser view attached).
The same issue was observed in internal PROseq samples as well.

Command used and terminal output

nextflow run nf-core/nascent -profile docker --input /home/ec2-user/environment/sampleSheets/sampleSheet.public_Jurkat.202306.csv \
    --assay_type PROseq \
    --tracedir ./pipeline_info \
    --fasta /home/ec2-user/environment/annotations/bwa/hg38as.fa \
    --bwa_index /home/ec2-user/environment/annotations/bwa \
    --gtf /home/ec2-user/environment/annotations/gencode.v38.chr_patch_hapl_scaff.annotation.gtf 
    --filter_bed /home/ec2-user/environment/annotations/hg38as.blacklist.ENCODEDCC.bed \
    -w /data/work --outdir /data/outdir/public.Jurkat.GSE66031/ -resume > public.Jurkat.202306.log 2>&1

Relevant files

image

System information

Nextflow version: 23.04.1
Hardware : AWS Cloud9
Executor : Local
Container engine : Docker
OS:
Nascent pipeline version: v2.1.1-g9ff33c7

Pass All CI

Description of the bug

The CI tests are all over the place currently.

Command used and terminal output

No response

Relevant files

No response

System information

No response

Refactor ext.when

Description of feature

Need to come up with some community guidelines for when these should be used.

ext.when = { params.save_reference }

Use config to control the configuration, not the execution logic.

Featurecounts only counts Exons

From @ranikay in slack

One question that we've run into when testing out the latest version on the dev branch - it appears that featureCounts is restricted to counting reads in exons. Does anyone know if that is intentional, and what best-practices would be in this area? It seems that intronic reads would be valid to count for PRO-seq.

BEDTOOLS_INTERSECT_FILTER is not invoked before accessing the output attribute in release version

Description of the bug

BEDTOOLS_INTERSECT_FILTER is not invoked before accessing the output attribute in script subworkflows/local/transcript_identification.nf at line 66 when params.filter_bed is not defined or set to false.

Fix:

remove codes from line 64 to 68 in file transcript_identification.nf

Command used and terminal output

No response

Relevant files

No response

System information

No response

GROHMM and CHM13

Description of the bug

grohmm fails with CHM13

Command used and terminal output

No response

Relevant files

No response

System information

No response

Example of 3D genome integration

Description of feature

Using 3D genome integration to show Enhancer-promoter pairs and what the targets are. Feedback from Michael Zhang.

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